Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Reexamination Certificate
1999-11-02
2001-12-04
Horlick, Kenneth R. (Department: 1656)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
C427S458000, C427S180000
Reexamination Certificate
active
06326149
ABSTRACT:
The present invention relates to methods of fabricating surfaces on which defined amounts of chemical components are present in spatially resolved segments, and to substrates fabricated with such defined, resolved segments on their surface.
The present invention pertains to improved fabrication methods and technology tools to increase the availability and diversity of low cost, point of care/home health testing kits. Such methods and technology tools include the dry deposition of charged reagents and powders, preferably medicaments, on a variety of surfaces for applications in the pharmaceutical industry and health care industry. These dry reagent deposition methods and tools can provide high speed, low cost manufacturing and thus are well suited to develop products for the home health testing industry, particularly for use in populations with limited access to low cost health care. Such products are user-friendly, non-mechanical, stable, accurate, precise, sensitive, specific and quantitative. The methods of the present invention are appropriate for using electrostatics to deposit biomolecule reagent microparticles on a membrane for a point of care/home health testing assay.
An important element of the present invention is the recognition that the amounts of biomolecules or the like that need to be associated with a substrate for useful diagnostic tools are often less than the amounts typically directly deposited by electrostatic deposition. These biomolecules can be associated with particles in an appropriate amount, and the particles are usefully employed in electrostatic depositions to economically and reliably fabricate test strips. Appropriate use of the deposition engine allows spatially resolved depositions of biomolecules, for instance to create reagents that develop visual indications such as “+” or “−” signs. Unexpectedly, the carrier particles and compositions used to retain the carrier particles do not interfere with bioassays.
SUMMARY OF THE INVENTION
The invention provides a method of fabricating a substrate having on a surface thereof two or more spatially-resolved regions, each with a defined amount or concentration of one of one or more chemical components, the method comprising: associating a chemical component with particles of 1 &mgr;m to 500 &mgr;m diameter; electrostatically depositing the particles on the appropriate regions; and if a said spatially-resolved region is to receive a second chemical component, repeating steps (a) and (b) for that chemical component and the appropriate regions. Preferably, the chemical component comprises 20% w/w or less (more preferably 10% w/w or less, or 5% w/w or less) of the particles In one embodiment, the chemical component is associated with preliminary particles, and the first particles are aggregated with other solid components to form the particles of 1 &mgr;m to 500 &mgr;m diameter. In certain embodiments, the chemical component is bioactive agent, a peptide, a protein, a nucleic acid, a nucleic acid-intercalating dye. The associating can, for example, comprise covalently attaching or associating with an association constant of at least 10
12
M
−l
. Preferably, the method further includes coating the regions with a polymer composition adapted to secure the particles to the surface.
The invention includes substrates produced by the method.
The invention further includes a method of fabricating a cell growth supporting substrate with spatially-resolved regions with associated bioactive agents comprising associating the bioactive agents with the cell growth supporting substrate or a separate substrate that is incorporated into the cell growth supporting substrate with the method. The bioactive agent can be, for example a nutrient or an antimicrobial agent.
The invention further includes a substrate comprising: a first layer comprising a chemical component supporting an assay reaction, the layer formed by the method; and a second layer comprising a second chemical component, distinct from the first, supporting the assay reaction. The substrate can comprise a reagent that facilitates visualization of a reaction product, such as colloidal gold or horse radish peroxidase.
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Bentz Bryan Lloyd
Chiang William
Loewy Zyi Gerald
Sun Hoi Cheong
Burke William J.
Horlick Kenneth R.
Sarnoff Corporation
Strzelecka Teresa
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